Novel endogenous promoters for genetic engineering of the marine microalga Nannochloropsis gaditana CCMP526

© 2019 Elsevier B.V. Nannochloropsis is a marine microalga from the Eustigmatophyceae stramenopile lineage that has been studied extensively due to a broad range of industrial applications, mostly related to their oil and pigment production. However, tools to genetically engineer members of this group, and therefore further understand and maximise their industrial potential are still limited. In order to expand the potential industrial uses of this organism, several molecular tools, including gene promoters of different strength, are needed. A comprehensive and diverse set of well-characterized promoters is key to a number of genetic engineering and synthetic biology applications, such as the assembly of complex biological functions or entire metabolic pathways. In this study, we measured the promoter activity of three endogenous constitutive promoters from N. gaditana genes EPPSII (Nga02101); HSP90 (Nga00934); ATPase (Nga06354.1) in driving the expression of a Sh ble- mVenus fluorescent reporter fusion protein. Through a combined approach that includes flow cytometry, RT-qPCR and immunoblotting, we profiled the activity of these promoters at both the transcript and protein level. Two promoters HSP90 (Nga00934) and EPPSII (Nga02101) outperformed the widely used β-tubulin promoter, exhibiting 4.5 and 3.1-fold higher mVenus fluorescence, respectively. A third promoter ATPase (Nga06354.1) was also able to drive the expression of transgenes, albeit at lower levels. We show that the new promoters identified in this study are valuable tools, which can be used for genetic engineering and functional genetics studies in N. gaditana

Doing synthetic biology with photosynthetic microorganisms

The use of photosynthetic microbes as synthetic biology hosts for the sustainable production of commodity chemicals and even fuels has received increasing attention over the last decade. The number of studies published, tools implemented, and resources made available for microalgae have increased beyond expectations during the last few years. However, the tools available for genetic engineering in these organisms still lag those available for the more commonly used heterotrophic host organisms. In this mini-review, we provide an overview of the photosynthetic microbes most commonly used in synthetic biology studies, namely cyanobacteria, chlorophytes, eustigmatophytes and diatoms. We provide basic information on the techniques and tools available for each model group of organisms, we outline the state-of-the-art, and we list the synthetic biology tools that have been successfully used. We specifically focus on the latest CRISPR developments, as we believe that precision editing and advanced genetic engineering tools will be pivotal to the advancement of the field. Finally, we discuss the relative strengths and weaknesses of each group of organisms and examine the challenges that need to be overcome to achieve their synthetic biology potential.Peer reviewe

The meaning of my feelings depends on who I am: work-related identifications shape emotion effects in organizations

Theory and research on affect in organizations has mostly approached emotions from a valence perspective, suggesting that positive emotions lead to positive outcomes and negative emotions to negative outcomes for organizations. We propose that cognition resulting from emotional experiences at work cannot be assumed based on emotion valence alone. Instead, building on appraisal theory and social identity theory, we propose that individual responses to discrete emotions in organizations are shaped by, and thus depend on, work-related identifications. We elaborate on this proposition specifically with respect to turnover intentions, theorizing how three discrete emotions - anger, guilt, and pride - differentially affect turnover intentions, depending on two work-related identifications - organizational and occupational identification. A longitudinal study involving 135 pilot instructors reporting emotions, work-related identifications, and turnover intentions over the course of one year provides general support for our proposition. Our theory and findings advance emotion and identity theories by explaining how the effects of emotions are dependent on the psychological context in which they are experienced

Construction of Stability Multipliers with Prescribed Phase Characteristics: an Improved Value for $\sigma

For a feedback system consisting of a transfer function $G(s)$ in the forward path and a time-varying gain $n(t)(0 \leqq n(t) \leqq k)$ in the feedback loop, a stability multiplier $Z(s)$ has been constructed (and used to prove stability) by Freedman [2] such that $Z(s)(G(s) + {1 / K})$ and $Z(s - \sigma )(0 < \sigma < \sigma _ * )$ are strictly positive real, where $\sigma _ *$ can be computed from a knowledge of the phase-angle characteristic of $G(i\omega ) + {1 / k}$ and the time-varying gain $n(t)$ is restricted by $\sigma _ *$ by means of an integral inequality. In this note it is shown that an improved value for $\sigma _ *$ is possible by making some modifications in his derivation. ©1973 Society for Industrial and Applied Mathematics

Synthesis and ¹H and ¹³C NMR spectral analysis of some spirolactones of 2,4-diaryl-3-azabicyclo [3.3.1]nonan-9-ones

96-100Introduction of an α-methylene-γ-butyrolactone unit at C-9 does not change the conformation of the bicyclononane ring system. However, NMR signals for H-1 and H-5 and axial H-7 protons are shifted upfield and the signals for H-2 and H-4 protons are shifted down field. The signals for all ring carbons are shifted upfield. Spectral data show that the C(9)C(10) bond of spirolactone is oriented towards the piperidine ring

L<SUB>2</SUB>-Stability of time-varying systems with global conditions on the time-varying gain

The L2-stability of linear und non-linoar time-varying systems is considered and integral typo condition on the time-varying gain is obtained through the positive operator theory